Insulin is a key component serum free growth media for mammalian cells, used for the manufacturing of monoclonal antibodies, virus vaccines, gene therapy products and other biological drugs. It stimulates the proliferation of cells. The yeast organism used to produce Human Insulin does not contain genes or express antigens of livestock or poultry disease agents, and is neither stored nor grown in media containing animal raw material. Human Insulin is considered to be virologically safe and the risk of transferring BSE agents to the media or reagents in which the recombinant human insulin is a component is negligible, since no animal derived raw material is used in the manufacturing process a recombinant enzyme is used in the process the production strain for manufacture is yeast, which is not the natural host for mammalian viruses
The recommended concentration of Insulin Human in cell culture media is 1-10/mg/L, depending on cell type and specific application.
Below are instructions for dilution of Human Insulin, Animal-free.
Suspend the insulin product in purified water.
Dissolve the insulin product by adjusting the pH to 3 with diluted HCl (0.1 or 0.01M, depending on the desired volume of growth medium). The insulin solution should not be held at pH 3 for more than one hour.
Readjust pH to 7.4 with buffer or base. The solution becomes unclear when passing through the isoelectric point of the insulin product, but becomes clear again as the pH of the solution reaches 7.
The solution should be sterile filtered using a low protein binding filter with a pore size of 0.2 µm. Aeration and foaming should be avoided during solubilization and filtration, as this can lead to protein denaturation.